Alternate Protocol 3: Production of Androgenetic Haploid Embryos

Materials

PVP360 solution: Polyvinylpyrrolidone (PVP360, average Mr≈360,000) at 5% to 20% (w/v; typically ∼10%) dissolved in sterile, high‐quality distilled water, and filter‐sterilized through
a 0.45‐μm filter (store in 1‐ml aliquots at −20°C, with one thawed for current use that may be kept at room temperature for
>1 month)

Air‐cushioned table or platform (suppliers respectively include Technical Manufacturing Corporation and Meiritsu); the need
for insulation against vibration depends on the location of the workstation; freezers, air‐conditioning, elevators and busy
nearby roads are common causes of persistent vibration

Piezo‐actuated micromanipulator, such as the PMAS‐CT150 (Prime Tech)

Mouth pipet for handling of embryos [this can be made from a Microcap bulb dispenser (Drummond Scientific Company) equipped
with an air‐tight assembly of silicone tubing attached to filtered blue Gilson tips and a pipet pulled by hand from 116‐mm
glass capillaries (Drummond, cat no. 1‐000‐1000)]